Cytokines And Angiogenic Factors Research Articles

581P - A biomarker combination indicating resistance to FOLFOX plus bevacizumab in metastatic colorectal cancer: Results of phase I of the PERMAD trial

BackgroundAntiangiogenic agents such as bevacizumab (bev) are widely used in combination with chemotherapy to treat metastatic colorectal cancer (mCRC). Predictive markers indicating resistance to antiangiogenic agents are elusive. Cytokines and angiogenic factors (CAF) may enable such a prediction. The PERMAD trial has two phases: Phase I, reported here, aims to establish a CAF marker combination (CAFmC) that enables early prediction of treatment resistance in treatment naïve patients with mCRC receiving bev plus mFOLFOX6. Phase II will prospectively evaluate this CAFmC and randomize between an early replacement of bev by aflibercept or continuation of bev when the CAFmC indicates imminent progress. MethodsIn phase I 41 out of 50 patients recruited in 15 centers in Germany and Austria with treatment naïve mCRC under FOLFOX plus bev treatment were evaluable for CAF analysis. 102 different, preselected CAFs were prospectively collected and centrally analyzed in plasma samples (n=647) obtained prior to treatment and biweekly until radiologic progress determined by CT scan every 2 months. The values of various CAFs were affected by both, chemotherapeutic treatment itself as well as progress. These CAF were excluded from the bioinformatic analysis. Using the remaining CAF we employed a machine learning approach to define a combination of 5 CAF whose change in values/pattern correlated with later progress at least 2 months prior to radiologic progress as determined by CT. Out of various classifiers examined, a random forest classifier provided a CAF set with the highest accuracy. ResultsUsing the samples described above and a random forest algorithm we established a CAFmC comprising 5 CAF whose specific change in value/pattern over time indicated treatment resistance 3 months prior to radiologic progress with an accuracy of 83%. The CAFmC was established and cross-validated in two cohorts of 26 and 15 patients, respectively. ConclusionsUsing advanced bioinformatics we identified a CAFmC that points out treatment resistance to FOLFOX plus Bev in patients with mCRC 3 months prior to radiological progress. A decision process using this marker combination will be evaluated in the randomized phase II of the trial. Clinical trial identificationNCT02331927, 2012-005657-24. Legal entity responsible for the studyUlm University Hospital. FundingSanofi-Aventis. DisclosureT. Seufferlein: Research grant / Funding (institution): Celgene; Research grant / Funding (institution): Sanofi; Advisory / Consultancy: Celgene; Advisory / Consultancy: Lilly; Advisory / Consultancy: Boeringer Ingelheim; Advisory / Consultancy: Merck Serono; Advisory / Consultancy: Amgen; Advisory / Consultancy: Roche. T.J. Ettrich: Advisory / Consultancy: Merck Serono; Advisory / Consultancy, Speaker Bureau / Expert testimony: Sanofi-Aventis; Advisory / Consultancy: Bayer; Advisory / Consultancy, Speaker Bureau / Expert testimony: Bristol-Myers Squibb; Advisory / Consultancy: Pfizer; Advisory / Consultancy: Novartis; Advisory / Consultancy: Roche; Advisory / Consultancy: Lilly; Speaker Bureau / Expert testimony: Celgene; Travel / Accommodation / Expenses: Ipsen; Research grant / Funding (institution): Servier. A. König: Travel / Accommodation / Expenses: Ipsen. L. Perkhofer: Travel / Accommodation / Expenses: Ipsen. All other authors have declared no conflicts of interest.

Prognostic or predictive value of circulating cytokines and angiogenic factors for initial treatment of multiple myeloma in the GIMEMA MM0305 randomized controlled trial

BackgroundSeveral new drugs are approved for treatment of patients with multiple myeloma (MM), but no validated biomarkers are available for the prediction of a clinical outcome. We aimed to establish whether pretreatment blood and bone marrow plasma concentrations of major cytokines and angiogenic factors (CAFs) of patients from a phase 3 trial of a MM treatment could have a prognostic and predictive value in terms of response to therapy and progression-free and overall survival and whether these patients could be stratified for their prognosis.MethodsBlood and bone marrow plasma levels of Ang-2, FGF-2, HGF, VEGF, PDGF-β, IL-8, TNF-α, TIMP-1, and TIMP-2 were determined at diagnosis in MM patients enrolled in the GIMEMA MM0305 randomized controlled trial by an enzyme-linked immunosorbent assay (ELISA). These levels were correlated both reciprocally and with the type of therapy and patients’ characteristics and with a group of non-MM patients as controls.ResultsNo significant differences were detected between the blood and bone marrow plasma levels of angiogenic cytokines. A cutoff for each CAF was established. The therapeutic response of patients with blood plasma levels of CAFs lower than the cutoff was better than the response of those with higher levels in terms of percentage of responding patients and quality of response.ConclusionFGF-2, HGF, VEGF, and PDGF-β plasma levels at diagnosis have predictive significance for response to treatment. The stratification of patients based on the levels of CAFs at diagnosis and their variations after therapy is useful to characterize different risk groups concerning outcome and response to therapy.Trial registrationClinical trial information can be found at the following link: NCT01063179

Plasma cytokine and angiogenic factors associated with prognosis and therapeutic response to sunitinib vs everolimus in advanced non-clear cell renal cell carcinoma.

No biomarkers are available to predict relative clinical benefit from targeted therapies in patients with non-clear cell renal cell carcinoma (nccRCC). To identify candidate predictive markers, we investigated a set of cytokines and angiogenic factors (CAFs) in previously untreated patients with nccRCC participating in the phase II ESPN trial comparing first-line sunitinib to everolimus. Pre-treatment concentrations of 30 CAFs were measured in plasma from 37 patients treated with everolimus (n=16) or sunitinib (n=21), and associated with progression-free (PFS) and overall survival (OS) after adjusting for potential confounders. High (>median) concentrations of soluble glycoprotein 130 (sgp130) were predictive of a longer PFS with sunitinib compared with everolimus (HR = 0.30; 95% CI: 0.11-0.85; P = 0.024). Significantly shorter PFS was noted, independently of treatment arm, in patients with high (>median) levels of IL-8 (HR = 3.13; 95% CI: 1.41-6.92), IL-13 (HR = 3.36; 95% CI: 1.49-7.58), and soluble tumor necrosis factor receptor II (HR = 2.21; 95% CI: 1.04-4.72). High IL-8 levels were also associated with significantly shorter OS (HR = 3.55; 95% CI: 1.55-8.14). Thus, using CAF profiling we identified candidate prognostic and predictive circulating biomarkers that can be used to inform therapeutic decisions in nccRCC.

Prognostic and predictive plasma cytokines and angiogenic factors (CAF) in patients (pts) with advanced non-clear cell renal cell carcinoma treated with first-line sunitinib or everolimus: Results from the ESPN trial.

e16112Background: There is a need for markers to guide treatment selection in non-clear cell renal cell carcinoma (nccRCC). We found that soluble tumor necrosis factor-α receptor-I (sTNFR-I) and in...

Prognostic implication of antitumor immunity measured by the neutrophil-lymphocyte ratio and serum cytokines and angiogenic factors in gastric cancer.

The neutrophil-lymphocyte ratio (NLR) is associated with a poor prognosis in many cancers but the biological mechanisms involved are unknown. Since cytokines and angiogenic factors (CAFs) are reflected by various immune responses, we analyzed the association between the NLR and CAFs and their prognostic implications in gastric cancer (GC). Of 745 GC patients who were enrolled in NLR analysis, 70 underwent NLR and CAF association analyses. Pretreatment serum levels of 52 CAFs were measured by means of multiplex bead immunoassays and enzyme-linked immunosorbent assays. Linear regression analysis and survival analysis of the NLR with each CAF were performed. Metastatic organ numbers and carbohydrate antigen 19-9 levels were significantly higher in patients with a high NLR [greater than 2.42 (median): P=0.047 and P<0.001 respectively]. The overall survival was significantly worse in the high NLR group (17.8months vs 11.2months, P<0.001). In CAF analysis, osteopontin (R 2=0.337, P<0.001) and interleukin-6 (R 2=0.141, P=0.001) were significantly associated with the NLR. Stromal-cell-derived factor 1 (SDF-1) was a significant poor prognostic factor independently of the NLR. Consideration of both the NLR and SDF-1 divided patient groups with different overall survival (both low, 21.0 months; either high, 15.8months; both high, 8.2months). The NLR is a significant poor prognostic factor in advanced GC. The NLR is mainly associated with osteopontin and interleukin-6. Besides the NLR, SDF-1 is an independent poor prognostic factor in GC. Consideration of both the NLR and SDF-1 might give insights into antitumor immunity in GC.

Dynamic Contrast-Enhanced Magnetic Resonance Imaging as a Surrogate Biomarker for Bevacizumab in Colorectal Cancer Liver Metastasis: A Single-Arm, Exploratory Trial.

PurposeThe purpose of this study is to investigate dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) and plasma cytokines and angiogenic factors (CAFs) as pharmacodynamic and prognostic biomarkers of bevacizumab monotherapy in colorectal cancer with liver metastasis (CRCLM).Materials and MethodsFrom July 2011 to March 2012, 28 patients with histologically confirmed CRCLM received bevacizumab monotherapy followed by combined FOLFOX therapy. The mean age of the patients was 57 years (range, 30 to 77 years). DCE-MRI (Ktransand IAUC60) was performed at baseline, first follow-up (3 days after bevacizumab monotherapy), and second follow-up (3 days after combined therapy). CAF levels (vascular endothelial growth factor [VEGF], placental growth factor [PlGF], and interleukin-8) were assessed on the same days. Progression-free survival (PFS) time distributions were summarized using the Kaplan-Meier method and compared using log-rank tests.ResultsThe median PFS period was 11.2 months. Ktrans, IAUC60, VEGF, and PlGF values on the first follow-up day were significantly different compared with baseline values. No differences were observed on the second follow-up day. A > 40% decrease in Ktrans from baseline to first follow-up was associated with a longer PFS (hazard ratio, 0.349; 95% confidence interval, 0.133 to 0.912; p=0.032). Changes in CAFs did not show correlation with PFS time.ConclusionDCE-MRI parameters and CAFs are pharmacodynamic biomarkers of bevacizumab for CRCLM. In our study, change in Ktrans at 3 days after bevacizumab monotherapy was a favorable prognostic factor; however, the value of CAFs as a prognostic biomarker was not found.

Signature of cytokines and angiogenic factors (CAFs) defines a clinically distinct subgroup of gastric cancer

Little is known about cytokine and angiogenic factors (CAFs) in gastric cancer (GC) in terms of tumor classification and prognostic value. Here, we aimed to correlate CAF signature with overall survival (OS) in GC. We measured pretreatment serum levels of 52 kinds of CAFs in 68 GC patients who were treated with fluoropyrimidine and platinum combination chemotherapy using multiplex bead immunoassays and enzyme-linked immunosorbent assay. We evaluated correlations between CAF levels and pathological features and OS. Three distinct patient groups were identified: one with high levels of proangiogenic factors, another with high levels of proinflammatory factors, and the other with high levels of both factors. Eleven CAFs [interleukin (IL)-2 receptor-alpha, growth-regulated alpha protein, hepatocyte growth factor, macrophage colony-stimulating factor, stromal cell-derived factor, IL-6, IL-8, IL-10, interferon-gamma, vascular endothelial growth factor, and osteopontin] were independently correlated with poor OS. Clustering analysis of these 11 CAFs revealed distinct high and low 11-CAF signature groups. High 11-CAF signature was associated with shorter OS (10.1 vs. 17.9months, p=0.026) along with poor performance status, and the presence of signet ring cell components in multivariate analysis of OS (HR 1.76, p=0.029). The patients' traditional clinicopathological characteristics were not significantly different between the high and low 11-CAF signature groups. Serum CAF profiling differentiated GC patient groups. A high 11-CAF signature could identify GC patients with a poor prognosis when treated with standard chemotherapy who need urgent new treatment strategies.

A plasma cytokine and angiogenic factor (CAF) analysis for selection of bevacizumab therapy in patients with metastatic colorectal cancer.

This study intends to identify biomarkers that could refine the selection of patients with metastatic colorectal cancer (mCRC) for bevacizumab treatment. Pretreatment 36 plasma cytokines and angiogenic factors (CAFs) were first measured by protein microarray analysis in patients who received first-line bevacizumab-containing therapies (discovery cohort, n = 64), and further evaluated by enzyme-linked immunosorbent assay in patients treated on regimens with or without bevacizumab (validation cohort, n = 186). Factor levels were correlated with clinical outcomes, predictive values were assessed using a treatment by marker interaction term in the Cox model. Patients with lower pretreatment levels of hepatocyte growth factor (HGF) or VEGF-A121 gain much more benefit from bevacizumab treatment as measured by progression-free survival (PFS) and overall survival (OS), while angiopoietin-like 4 (ANGPTL4) levels negatively correlated with PFS and response rate following bevacizumab (all adjusted interaction P < 0.05). A baseline CAF signature combining these three markers has greater predictive ability than individual markers. Signature-negative patients showed impaired survival following bevacizumab treatment (PFS, 7.3 vs 7.0 months; hazard ratio [HR] 1.03; OS, 29.9 vs 21.1 months, HR 1.33) compared with signature-positive patients (PFS, 6.5 vs 11.9 months, HR 0.52; OS, 28.0 vs 55.3 months, HR 0.67). These promising results warrant further prospective studies.

A cytokine and angiogenic factor (CAF) analysis in plasma in testicular germ cell tumor patients (TGCTs).

A cytokine and angiogenic factor (CAF) analysis in plasma in testicular germ cell tumor patients (TGCTs).

Abstract 29: Myeloid cell and plasma cytokine and angiogenic factor (CAF) dynamics in association with response to VEGFR inhibitors cediranib and sunitinib

Abstract Background: Anti-angiogenic agents targeting the VEGF pathway have improved the outcomes of patients with metastatic renal cell carcinoma (mRCC), but therapy resistance consistently occurs. In RCC, the presence of increased intratumoral and peripheral blood granulocytes is associated with decreased survival. Neutrophilic and monocytic forms of myeloid cells have been proposed to contribute to immune evasion, metastasis, and anti-VEGF therapy resistance by secreting pro-angiogenic and pro-tumorigenic factors. We sought to evaluate changes in these cells and CAFs in determining response to VEGFR tyrosine kinase inhibitors (TKis). Methods: We analyzed the time-course effects of VEGFR TKis cediranib (10 mg/kg/day) and sunitinib (50 mg/kg/day) on myeloid cells and CAFs in partially treatment sensitive syngeneic mouse models of RCC (RenCa, N=10) and breast cancer (4T1, N=10). Peripheral blood, subcutaneous tumors, and spleens were collected at baseline before treatment, at maximum response, and progression. We evaluated changes in macrophages (F4/80+), monocytic (Ly6C+Ly6G low) and neutrophilic (Ly6C low Ly6G+) populations of CD11b+ myeloid cells and VEGFR1+ cells in the spleens and tumors. Finally, we profiled 144 cytokines, angiogenic factors to characterize patterns of expression and correlate them with changes in myeloid cell populations and dynamics of response. Results: Cediranib or sunitinib treatment arrested tumor growth in both RenCa and 4T1, yet 4T1 tumors grew back on treatment earlier on time. We found a trend of increase in monocytic CD11b+ cells in the tumors during VEGFR TKi treatment, highest at the time of progression, and a less clear decrease in the neutrophilic subset. Intratumoral F4/80+ macrophages decreased with cediranib/sunitinib, and showed a maximum decline at the time of progression. Changes in immune cells particularly in 4T1 tumors were accompanied by accumulation of neutrophilic CD11b+ cells in the spleen at the time of progression; however, the splenic monocytic CD11b+ cells decreased on treatment and remained suppressed. Preliminary CAF analysis revealed a dramatic elevation of G-CSF particularly in 4T1 mice, while GM-CSF levels remained steady. Conclusions: Putative pro-tumorigenic monocytic CD11b+ myeloid cells increase during treatment with cediranib/sunitinib in partially sensitive RenCa and 4T1 tumors, suggesting a role in resistance. The splenic accumulation of the neutrophilic subset at the time of progression did not correlate with a similar presence in the tumors. Integrative analysis of myeloid cell population and plasma CAF dynamics may provide relevant information to better understand VEGFR TKi resistance and will be shown at the meeting. Citation Format: Alper Yetil, John V. Heymach, Amado J. Zurita. Myeloid cell and plasma cytokine and angiogenic factor (CAF) dynamics in association with response to VEGFR inhibitors cediranib and sunitinib. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 29. doi:10.1158/1538-7445.AM2014-29

Correlation between circulating tumor cells (CTC) and plasma cytokines and angiogenic factors (CAF) in early breast cancer patients.

e22038 Background: CAFs are involved in cancer invasion and metastasis. CTCs play role in tumor dissemination and are an independent survival predictor in breast cancer (BC) patients. The aim of th...

Modulation of Circulating Angiogenic Factors and Tumor Biology by Aerobic Training in Breast Cancer Patients Receiving Neoadjuvant Chemotherapy

Aerobic exercise training (AET) is an effective adjunct therapy to attenuate the adverse side-effects of adjuvant chemotherapy in women with early breast cancer. Whether AET interacts with the antitumor efficacy of chemotherapy has received scant attention. We carried out a pilot study to explore the effects of AET in combination with neoadjuvant doxorubicin-cyclophosphamide (AC+AET), relative to AC alone, on: (i) host physiology [exercise capacity (VO2 peak), brachial artery flow-mediated dilation (BA-FMD)], (ii) host-related circulating factors [circulating endothelial progenitor cells (CEP) cytokines and angiogenic factors (CAF)], and (iii) tumor phenotype [tumor blood flow ((15)O-water PET), tissue markers (hypoxia and proliferation), and gene expression] in 20 women with operable breast cancer. AET consisted of three supervised cycle ergometry sessions/week at 60% to 100% of VO2 peak, 30 to 45 min/session, for 12 weeks. There was significant time × group interactions for VO2 peak and BA-FMD, favoring the AC+AET group (P < 0.001 and P = 0.07, respectively). These changes were accompanied by significant time × group interactions in CEPs and select CAFs [placenta growth factor, interleukin (IL)-1β, and IL-2], also favoring the AC+AET group (P < 0.05). (15)O-water positron emission tomography (PET) imaging revealed a 38% decrease in tumor blood flow in the AC+AET group. There were no differences in any tumor tissue markers (P > 0.05). Whole-genome microarray tumor analysis revealed significant differential modulation of 57 pathways (P < 0.01), including many that converge on NF-κB. Data from this exploratory study provide initial evidence that AET can modulate several host- and tumor-related pathways during standard chemotherapy. The biologic and clinical implications remain to be determined.

A cytokine and angiogenic factor (CAF) analysis in plasma in testicular germ cell tumor patients (TGCTs).

e15599 Background: We investigated cytokines and angiogenic factors (CAFs) in patients with testicular germ cell tumors (TGCTs). We aimed to link the CAF profile to type of response to chemotherapy and select candidate prognostic and predictive markers for further study. Methods: In the presented study, plasma from 99 patients (pt) with TGCTs treated with first line (95 pt) or salvage (4 pt) chemotherapy was collected. The concentrations of 51 plasma CAFs were measured pre-treatment (n = 80) and on day 22 (n = 60) using multiplex bead arrays (Human Group I and II cytokine panels and TGF beta by Bio-Plex 200 system (Bio-Rad Laboratories, Hercules, CA). We used unsupervised clustering with self-organizing map (SOM) and k-means clustering to analyze CAF expression profiles. Results: Unsupervised clustering with self-organizing maps and k-means identified characteristic plasma cytokine profiles in different subgroups of patients according to response to chemotherapy and other clinical variables. Several cytokines were differentially expressed in patients with favourable and unfavorable response in serum before chemotherapy including IL-1b, IL-15, M-CSF, IL-4, IL-5, b-NGF, IL-10, MCP-3, FGF basic, IL-6, MIP-1a, GM-CSF, IL-17, IL-13, MIP1b, IL-8, SCF, IFN alfa, SCGFb, IL-2RA, IP-10, IL-16, TGFb-3. Similarly, following cytokines were differentially expressed in serum after 1st cycle of chemotherapy IL-1b, TGFb-3, LIF, TGFb-2, IL-16, IL-18, IL-2RA, MCP-3, IFN alfa, HGF, IL1a, MIF, SCF, IL-3, SCGFb, b-NGF, MIG, CTACK. Conclusions: CAF profiling with unsupervised clustering revealed clinically relevant differences in subgroups of TGCTs patients. We suggest that this platform may provide valuable insights into TGCTs biology, and could help to identify plasma cytokine signature for predicting treatment resistance.

Serum insulin to predict time to castration-resistant progression and overall survival in metastatic androgen-dependent prostate cancer (mADPCa).

e16038 Background: Duration of response to androgen-deprivation therapy (ADT) is highly variable in patients with mADPC and prognostic markers are needed. Insulin resistance and hyperinsulinemia may contribute to prostate cancer progression. We hypothesized that pretreatment serum insulin levels would predict time to castration-resistant progression (PFS) and overall survival (OS). Methods: Sera from men treated on a randomized phase 3 trial of first line ADT vs. ADT plus chemotherapy were retrospectively analyzed using a multiplex ELISA for cytokines and angiogenic factors (CAFs). Univariate and multivariate Cox proportional hazards regression models were used to identify associations between CAFs and PFS/OS. Results: 66 pts were evaluable, 86% Caucasian, median age 72 yrs, median PSA 31.5ng/mL, 77% Gleason score of ≥8, and 53% high volume metastatic disease (HVM). Thirty-five pts received ADT; 31 pts received ADT+chemo. In univariate analysis, higher pretreatment insulin and C-peptide were positively correlated with PFS, whereas higher hepatocyte-growth factor (HGF), osteopontin (OPN) and HVM were negatively correlated with PFS. In multivariate analysis, only higher insulin was associated with longer PFS (HR=0.72, 95%CI 1.32 -0.87; p&lt;0.001), whereas higher HGF and OPN were associated with reduced PFS (HR=1.82, 95%CI 0.59-2.83, p&lt;0.01 and HR=1.81, 95%CI 1.18-2.47, p&lt;0.001, respectively). Higher Insulin and Program Death 1 (PD1) were associated with longer OS on multivariate analysis (HR=0.78 p&lt;0.02 and HR=0.55 p&lt;0.02, respectively), whereas HVM and higher OPN were associated with reduced OS (HR=2.28 p&lt;0.01 and HR=1.60 p&lt;0.02). Using low insulin, high HGF and high OPN as 3 independent risk factors (RF), 3 distinct risk groups could predict PFS: good (zero RF), intermediate (1 or 2 RF) and poor risk (3 RF), with median PFS of 6.90, 1.97, and 0.86 years, respectively (p&lt;0.001). Conclusions: Higher pretreatment insulin was associated with prolonged PFS and OS in men with mADPC treated with ADT. Our data suggest that insulin levels are a biomarker for sensitivity to ADT and highlight the complex interactions between metabolism and PCa progression.

Abstract 4645: Plasma cytokine and angiogenic factor (CAF) profiling for identification of markers associated with clinical benefit from bevacizumab (BEV) and with primary tumor presence in a phase II presurgical study of metastatic renal cell carcinoma (mRCC) patients

Abstract Purpose: The main objectives of this exploratory analysis were to identify biomarkers in blood associated with clinical benefit from bevacizumab, and to assess CAF changes related to nephrectomy (Nx) in mRCC. Methods: Plasma samples were obtained from 44 pts participating in a frontline presurgical study of bevacizumab in clear cell mRCC (22 pts treated with BEV and&amp;#x2028; 22 pts with BEV and erlotinib) (Jonasch et al. J Clin Oncol 2009), before treatment (BL), after 8 weeks of therapy/pre-nephrectomy (Nx), after 16 weeks of therapy/postNx, and every 4 weeks thereafter. Multiplex bead suspension arrays and ELISA were used to measure concentrations of 58 CAFs including VEGF, VEGF-C, placental growth factor (PlGF), soluble CAIX (sCA9), hepatocyte growth factor (HGF), osteopontin (OPN), matrix metalloproteinase-9 (MMP9), transforming growth factor (TGF)b1, endoglin, type IV collagen (ColIV), and multiple chemokines and interleukins (IL). CAFs were correlated with progression-free survival (PFS), overall survival (OS), and best response in primary and metastatic disease (% relative to BL). Results: Of 58 CAFs available at BL, high (above median) concentrations of TRAIL and ColIV were associated with longer PFS (7.6 v 2.8 and 8.1 v 2.9 months [mos], respectively; both P=0.001) and OS (27.9 v 13.2 and 32.8 v 14.3 mos, respectively; P=0.023 and P=0.025). In addition to high TRAIL, low levels of VEGF and PlGF also showed a significant association with enhanced response both in primary disease and metastases (all P&amp;lt;0.05). The combination BEV+erlotinib resulted in differential modulation of several CAFs relative to BEV from BL to preNx, including E-selectin, TRAIL and stem cell factor (SCF). Multiple CAFs, most prominently sCA9, endoglin, OPN, VEGF, HGF, and E-selectin, significantly decreased postNx, while TRAIL, SCF, and CTACK increased (relative to BL and also preNx; all P&amp;lt;0.05). Conclusions: We identified TRAIL, ColIV, VEGF and PlGF as candidate CAFs associated with clinical benefit from BEV in clear cell mRCC. The combination of BEV with erlotinib resulted in distinct on treatment CAF changes. The integration of these data with CAF profiling studies in pts with non-metastatic RCC could help define CAF sets useful to assist in risk stratification of RCC pts and provide insights into the biology of RCC progression. Citation Format: Alper Yetil, Eric Jonasch, Xuemei Wang, Christopher Wood, Surena Matin, Hai T. Tran, Nizar M. Tannir, John V. Heymach, Amado J. Zurita. Plasma cytokine and angiogenic factor (CAF) profiling for identification of markers associated with clinical benefit from bevacizumab (BEV) and with primary tumor presence in a phase II presurgical study of metastatic renal cell carcinoma (mRCC) patients [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4645. doi:10.1158/1538-7445.AM2013-4645

865P - A Cytokine and Angiogenic Factor (CAF) Analysis in Plasma in Testicular Germ Cell Tumor Patients

ABSTRACT Background Testicular germ-cell tumours (TGCTs) represent a model for the cure of cancer. Nonetheless, a small proportion of patients develop disease recurrence. We investigated cytokines and angiogenic factors (CAFs) in patients with TGCTs. We aimed to link the CAF profile to PFS and select candidate predictive and prognostic markers for further study. Methods In this ongoing translational study, plasma from 55 patients with TGCTs was collected. The concentrations of 51 plasma CAFs were measured pretreatment (n = 47) and on day 22 (n = 30) using multiplex bead arrays (Human Group I and II cytokine panels and TGF beta by Bio-Plex 200 system (Bio-Rad Laboratories, Hercules, CA). We investigated the association between baseline levels of CAFs and clinico-pathological variables. Results We observed elevated level of transforming growths factor beta1 (TGF-b1), IL-2Ra, CXCL9, CXCL10, IFN-gama, macrophage inflammatory protein-1b (MIP-1b), and platelet-derived growth factor-BB (PDGF-BB) in seminoma patients compared to non-seminoma. Patients with poor prognosis according to IGCCCG have elevated pretreatment level of beta-nerve growth factor (NGF-b) and stromal cell-derived factor-1 (SDF-1a) compared to patients with good/intermediate prognosis. Patients with metastatic disease had elevated pretreatment level of stem cell growth factor beta (SCGF-b) and PDGF-BB compared to patients with stage I disease. Conclusions Using CAF profiling, we revealed differences in subgroups of TGCTs patients. We suggest that this platform may provide valuable insights into TGCTs biology, and could be useful in identification of new therapeutic targets. Disclosure All authors have declared no conflicts of interest.

791PD - Multivariate Analysis of Cytokines and Angiogenic Factors (CAFS) and Established Prognostic Parameters in Metastatic Renal Cell Carcinoma (MRCC) Patients (PTS) Receiving Pazopanib or Placebo

ABSTRACT Background In mRCC, prognosis is still solely determined based on clinical criteria. Although multiple candidate biomarkers exist, none has yet been incorporated into practice. We identified CAFs associated with PFS in mRCC pts with ECOG PS £1 treated in a randomized, placebo-controlled, phase III clinical trial of pazopanib (Sternberg, JCO 2010, Tran, ASCO 2010 #4522). In this study, we evaluated the prognostic significance of these CAFs relative to established clinical parameters. Methods Seven candidate CAFs (IL-6, IL-8, VEGF, HGF, TIMP1, osteopontin [OPN], E-Selectin) in plasma obtained pretreatment, and 5 clinical variables indicative of poor prognosis (time from diagnosis to treatment upper [U]LN [Heng, JCO 2009]) were correlated with PFS (n = 343; n = 225 pazopanib/n = 118 placebo) using uni- and multivariable Cox proportional hazard models. Correlations with ECOG PS, Motzer's (MSKCC) and Heng's risk criteria were also assessed. CAFs were measured using SearchLight (Aushon). Results Median PFS for all pts was 29.9 weeks (95% CI 24.6–32.1), 39.6 weeks (95% CI 32.1-48.1) for those treated with pazopanib and 13.4 weeks (95% CI 12.1–19.1) for placebo. IL-6, IL-8, and OPN were prognostic [Table]. Considering treatment group, 2 clinical variables (hemoglobin ULN [HR 1.95, p = 0.0151]) were predictors of short PFS in multivariate analysis. Adjusting for those 2, only OPN and IL-6 in the placebo arm (OPN high 7.9 v low 24 weeks, p = 0.034; IL-6 high 7.9 v low 24.1 weeks, p = 0.03), and OPN alone in the pazopanib arm (high 31.3 v low 56.4 weeks, p = 0.0007), remained prognostic. Although the 3 clinical classifications were associated with PFS, none was as strong a prognostic marker as the CAFs [Table]. Conclusion In mRCC pts with good PS, circulating OPN and IL-6 are prognostic for PFS independent of established clinical criteria. CAF PFS (wks) P Clinical PFS (wks) P IL-6 Low 24·0 ECOG 0 18·4 0·144 High 9·9 1 13·0 OPN Low 24·0 MSKCC Good 24·0 0·011 High 8·4 Interm/Poor 12·1 IL-8 Low 23·9 0·002 Heng Good 24·3 0·139 High 8·4 Interm/Poor 12·6 Disclosure Y. Liu: Stock ownership: Yes; GSK Membership on an advisory board or board of directors: No Corporate-sponsored research: No Other substantive relationships: No . L.N. Pandite: Stock ownership: Yes; GSK Membership on an advisory board or board of directors: No Corporate-sponsored research: No Other substantive relationships: No. J.V. Heymach: Stock ownership: No Membership on an advisory board or board of directors: Yes; GSK Corporate-sponsored research: Yes; GSK for biomarker analysis Other substantive relationships: No. All other authors have declared no conflicts of interest.

Cytokine and angiogenic factors associated with efficacy and toxicity of pazopanib in advanced soft-tissue sarcoma: an EORTC-STBSG study

Background:Pazopanib has activity in relapsed non-adipocytic soft-tissue sarcomas (STS). A series of serum cytokines and angiogenic factors (CAFs) at baseline and changes in soluble vascular endothelial growth factor receptor-2 (sVEGFR2) or placental-derived growth factor (PlGF) levels during treatment were explored for their association with outcome.Methods:Twenty-three baseline CAFs, and sVEGFR2 and PlGF changes were measured in 85 and 32 patients, respectively. Associations between baseline CAF levels and efficacy parameters, plus between-week 12 sVEGFR2 and PlGF levels and pazopanib-specific toxicities were investigated.Results:At baseline, low interleukin (IL)-12 p40 subunit and MPC3 levels were associated with better progression-free survival (PFS) at 12 weeks (PFS12wks), low basic nerve growth factor and hepatocyte growth factor with a better PFS, and low inter-cellular adhesion molecule-1 and IL-2 receptor alpha with prolonged overall survival (OS; all P<0.05). Pazopanib decreased sVEGFR2 and increased PlGF levels. Low sVEGFR2 and high PlGF levels at week 12 were associated with higher-grade hypertension, with TSH elevations and with poorer PFS12wks, and OS (both P<0.05).Conclusion:Several baseline CAFs were related to outcome parameters. Low sVEGFR2 and high PlGF at week 12 associate with several pazopanib-specific toxicities and poorer efficacy. If confirmed, these factors may be used as early markers for response to and toxicity from pazopanib, enabling further individualisation of STS treatment.

Efficacy, Safety, and Biomarkers of Single-Agent Bevacizumab Therapy in Patients with Advanced Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC) is a highly vascularized tumor in which neoangiogenesis contributes to growth and metastasis. We assessed the safety, efficacy, and potential biomarkers of activity of bevacizumab in patients with advanced HCC. In this phase II trial, eligible patients received bevacizumab, 5 mg/kg or 10 mg/kg every 2 weeks. The disease-control rate at 16 weeks (16W-DCR) was the primary endpoint. Circulating endothelial cells (CECs) and plasma cytokines and angiogenic factors (CAFs) were measured at baseline and throughout treatment. The 16W-DCR was 42% (95% confidence interval, 27%-57%). Six of the 43 patients who received bevacizumab achieved a partial response (objective response rate [ORR], 14%). Grade 3-4 asthenia, hemorrhage, and aminotransferase elevation occurred in five (12%), three (7%), and three (7%) patients, respectively. During treatment, placental growth factor markedly increased, whereas vascular endothelial growth factor (VEGF)-A dramatically decreased (p < .0001); soluble VEGF receptor-2 (p < .0001) and CECs (p = .03) transiently increased on day 3. High and increased CEC counts at day 15 were associated with the ORR (p = .04) and the 16W-DCR (p = .02), respectively. Lower interleukin (IL)-8 levels at baseline (p = .01) and throughout treatment (p ≤ .04) were associated with the 16W-DCR. High baseline IL-8 and IL-6 levels predicted shorter progression-free and overall survival times (p ≤ .04). Bevacizumab is active and well tolerated in patients with advanced HCC. The clinical value of CECs, IL-6, and IL-8 warrants further investigation.

Plasma cytokines and angiogenic factors (CAFs) for evaluation of risk of recurrence and classification of patients (pts) with nonmetastatic renal cell carcinoma (RCC).

e15075 Background: Although recent years have brought great progress into understanding the pathogenesis of RCC and multiple promising biomarkers, no tumor tissue or blood-based test has yet been established to assist in the clinical determination of prognosis. Whether CAFs can be used to classify RCC pts and the set of CAFs that is host vs. tumor-dependent remain unclear. The main objectives of this exploratory analysis were to identify CAFs predictive of recurrence, and to gain insight into coordinated regulation and tumor dependency of individual factors by comparing CAFs pre- and post-nephrectomy (Nx) in RCC pts. Methods: Plasma samples were obtained before and up to 1 year after Nx from 52 pts with sporadic, non-metastatic RCC with dominant clear cell histology. Pts were categorized based on pathologic staging (pT1/pT2 as low-risk [LR] for recurrence, n=32; pT3/pT4 as high-risk [HR], n=20). Multiplex bead suspension arrays and ELISA were used to measure concentrations of 58 CAFs including angiogenesis and invasion mediators (VEGF, VEGF-C, soluble CAIX [sCA9], HGF and TGF-b1, among others), and multiple inflammatory and/or immunomodulatory chemokines and interleukins (IL). t-test and Wilcoxon signed rank or rank-sum tests were respectively used to assess changes in CAFs between pre- and post-Nx or to compare CAFs between subgroups of pts. Results: The concentrations of IL-8 and sCA9, among others, were significantly higher in HR vs. LR RCC pts pre-Nx (P&lt;0.01). The pre-Nx concentrations of e-selectin were lower in RCC pts with eventual disease recurrence (P=0.027). Several CAFs decreased significantly post-Nx, including sCA9 and endoglin, whereas others increased, such as stem cell factor, IL-12, IL-18 and vascular cell adhesion molecule-1. Angiogenic and hypoxia-regulated CAFs such as OPN, sCA9, HGF, endoglin and collagen type IV grouped together using unsupervised hierarchical clustering. Conclusions: Pre-Nx CAFs associated with risk for recurrence could be useful in risk stratification of non-metastatic clear cell RCC pts. Some tumor-dependent angiogenic CAFs are coordinately regulated in these pts.